Search: onr:"swepub:oai:DiVA.org:su-113574" >
A Cell-Free Translo...
A Cell-Free Translocation System Using Extracts of Cultured Insect Cells to Yield Functional Membrane Proteins
-
Ezure, Toru (author)
-
Nanatani, Kei (author)
-
Sato, Yoko (author)
-
show more...
-
Suzuki, Satomi (author)
-
Aizawa, Keishi (author)
-
Souma, Satoshi (author)
-
Ito, Masaaki (author)
-
Hohsaka, Takahiro (author)
-
- von Heijine, Gunnar (author)
- Stockholms universitet,Institutionen för biokemi och biofysik
-
Utsumi, Toshihiko (author)
-
Abe, Keietsu (author)
-
Ando, Eiji (author)
-
Uozumi, Nobuyuki (author)
-
show less...
-
(creator_code:org_t)
- 2014-12-08
- 2014
- English.
-
In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 9:12, s. e112874-
- Related links:
-
https://doi.org/10.1...
-
show more...
-
https://journals.plo...
-
https://urn.kb.se/re...
-
https://doi.org/10.1...
-
show less...
Abstract
Subject headings
Close
- Cell-free protein synthesis is a powerful method to explore the structure and function of membrane proteins and to analyze the targeting and translocation of proteins across the ER membrane. Developing a cell-free system based on cultured cells for the synthesis of membrane proteins could provide a highly reproducible alternative to the use of tissues from living animals. We isolated Sf21 microsomes from cultured insect cells by a simplified isolation procedure and evaluated the performance of the translocation system in combination with a cell-free translation system originating from the same source. The isolated microsomes contained the basic translocation machinery for polytopic membrane proteins including SRP-dependent targeting components, translocation channel (translocon)-dependent translocation, and the apparatus for signal peptide cleavage and N-linked glycosylation. A transporter protein synthesized with the cell-free system could be functionally reconstituted into a lipid bilayer. In addition, single and double labeling with non-natural amino acids could be achieved at both the lumen side and the cytosolic side in this system. Moreover, tail-anchored proteins, which are post-translationally integrated by the guided entry of tail-anchored proteins (GET) machinery, were inserted correctly into the microsomes. These results showed that the newly developed cell-free translocation system derived from cultured insect cells is a practical tool for the biogenesis of properly folded polytopic membrane proteins as well as tail-anchored proteins.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
Find in a library
-
PLOS ONE
(Search for host publication in LIBRIS)
To the university's database
- By the author/editor
-
Ezure, Toru
-
Nanatani, Kei
-
Sato, Yoko
-
Suzuki, Satomi
-
Aizawa, Keishi
-
Souma, Satoshi
-
show more...
-
Ito, Masaaki
-
Hohsaka, Takahir ...
-
von Heijine, Gun ...
-
Utsumi, Toshihik ...
-
Abe, Keietsu
-
Ando, Eiji
-
Uozumi, Nobuyuki
-
show less...
- About the subject
-
- NATURAL SCIENCES
-
NATURAL SCIENCES
-
and Biological Scien ...
- Articles in the publication
-
PLOS ONE
- By the university
-
Stockholm University